Combinations for the treatment of immunoinflammatory disorders and proliferative skin diseases

ABSTRACT

The invention features a method for treating a patient who has an immunoinflammatory disorder or a proliferative skin disease, or is at risk for developing an immunoinflammatory disorder or a proliferative skin disease, by administering to the patient a prostaglandin and a retinoid simultaneously or within 14 days of each other in amounts sufficient to reduce or inhibit immunoinflammatory or dermal/epidermal proliferation.

BACKGROUND OF THE INVENTION

The invention relates to the treatment of immunoinflammatory disordersand proliferative skin diseases.

Immunoinflammatory disorders (e.g., rheumatoid arthritis, psoriasis,ulcerative colitis, Crohn's disease, stroke-induced brain cell death,septic shock syndrome, ankylosing spondylitis, fibromyalgia,inflammatory dermatoses, asthma, multiple sclerosis, type I diabetes,systemic lupus erythematosus, scleroderma, systemic sclerosis, andSjögren's syndrome) are characterized by dysregulation of the immunesystem and inappropriate activation of body's defenses, resulting indamage to healthy tissue.

One percent of humans world-wide are afflicted with rheumatoidarthritis, a relentless, progressive disease causing severe swelling,pain, and eventual deformity and destruction of joints. According to theArthritis Foundation, rheumatoid arthritis currently affects over twomillion Americans, of which women are three times more likely to beafflicted. Rheumatoid arthritis is characterized by inflammation of thelining of the joints and/or other internal organs, and the presence ofelevated numbers of lymphocytes and high levels of proinflammatorycytokines.

Treatment of rheumatoid arthritis generally includes administration of(i) non-steroidal anti-inflammatories (e.g., detoprofen, diclofenac,diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen, indomethacin,ketoprofen, meclofenameate, mefenamic acid, meloxicam, nabumeone,naproxen sodium, oxaprozin, piroxicam, sulindac, tolmetin, celecoxib,rofecoxib, aspirin, choline salicylate, salsalte, and sodium andmagnesium salicylate); (ii) steroids, (e.g., cortisone, dexamethasone,hydrocortisone, methylprednisolone, prednisolone, prednisone,triamcinolone); (iii) DMARDs, i.e., disease modifying antirheumaticdrugs, (e.g., cyclosporine, azathioprine, methotrexate, leflunomide,cyclophosphamide, hydroxychoroquine, sulfasalazine, D-penicillamine,minocycline, and gold); or (iv) recombinant proteins (e.g., entanercept;soluble TNF receptor) and remicade (infliximab)).

Psoriasis is a common chronic proliferative skin disease, affecting upto 2% of the population. One characteristic of psoriasis is a stronghyperproliferation of epidermal keratinocytes and an incompleteepidermal differentiation that leads to severe scaling of the affectedskin areas. This proliferative event is accompanied by an inflammationof the epidermis and dermis, with infiltrates of T-cells, neutrophils,and macrophages. Consequently, psoriasis has characteristics of both anautoimmune disease and a proliferative skin disease.

SUMMARY OF THE INVENTION

We have discovered that the combination of a prostaglandin, alprostadil(also known as prostaglandin E1; (11α, 13E,15S)-11,15-dihydroxy-9-oxoprost-13-enoic acid; 11α,15α-dihydroxy-9-oxo-13-trans-prostenoic acid; or3-hydroxy-2-(3-hydroxy-1-octenyl)-5-oxocyclopentaneheptanoic acid), anda retinoid, tretinoin (also known as vitamin A; all trans retinoic acid;or3,7-dimethyl-9-(2,6,6-trimethylcyclohex-1-enyl)nona-2,4,6,8-all-trans-tetraenoicacid), brings about substantial suppression of TNFα levels induced inwhite blood cells. TNFα is a major mediator of inflammation. Specificblockade of TNF using antibodies or soluble receptors is a potenttreatment for patients having an immunoinflammatory disease, such asinflammatory bowel disease or rheumatoid arthritis, or a proliferativeskin disease. Thus, this combination can be used to treatimmunoinflammatory disorders and proliferative skin diseases. Moreover,based on the shared action among prostaglandin family members and amongretinoid family members, alprostidil and/or tretinoin can be replaced bya family member in the combination.

Accordingly, the invention features a method for treating a patient whohas, or is at risk for developing, an immunoinflammatory disorder orproliferative skin disease, by administering to the patient (i) aprostaglandin; and (ii) a retinoid, in amounts that treat the patient.The prostaglandin and the retinoid may be administered separately or ascomponents of a pharmaceutical composition.

The prostaglandin and retinoid can be administered within ten days ofeach other (e.g, within five days, twenty-four hours, or one hour ofeach other, or even simultaneously). Administration of each compound canoccur 1-4 times each day, or as necessary to alleviate symptoms.

The specific amounts of prostaglandin and retinoid administered dependon the specific combination of components (i.e., the specificprostaglandin/retinoid combination) and the mode of administration.Generally, when orally administered, the prostaglandin is administeredat a dose of 1 pg to 10 mg per day, desirably 10 pg to 1 mg per day,more desirably 1 to 500 μg per day, and most desirably 10 to 100 μg perday, while the retinoid is administered at a dose of 1 μg to 5 g perday, desirably 0.1 mg to 1 g per day, more desirably 1 to 100 mg perday, and most desirably 5 to 50 mg per day.

Generally, when administered by intravenous, intramuscular, orsubcutaneous injection, the prostaglandin is administered at a dosage of1 pg to 10 mg per day, desirably 10 pg to 1 mg per day, more desirably 1to 500 μg per day, and most desirably 10 to 100 μg per day, while theretinoid is administered at a dosage of 1 μg to 5 g per day, desirably0.1 mg to 1 g per day, more desirably 1 to 100 mg per day, and mostdesirably 5 to 50 mg per day.

Generally, when delivered by topical, transdermal, or ophthalmicapplication, or inhalation, rectal, or vaginal administration, theprostaglandin is administered at a dose of 1 pg to 100 mg per day,desirably 10 pg to 10 mg per day, more desirably 100 pg to 1 mg per day,and most desirably 0.01 to 0.5 mg per day, while the retinoid isadministered at a dose of 50 ng to 500 mg per day, desirably 500 ng to50 mg per day, more desirably 5 μg to 5 mg per day, and most desirably50 to 500 μg per day.

The invention also features a method for identifying compounds usefulfor treating a patient having an immunoinflammatory disorder or aproliferative skin disease. The method includes the steps of: contactingimmune cells in vitro with (i) a prostaglandin or a retinoid; and (ii) acandidate compound, and determining whether the immune response ismodulated relative to (a) immune cells contacted with the prostaglandinor retinoid but not contacted with the candidate compound, and (b)immune cells contacted with the candidate compound but not with theprostaglandin or retinoid. A candidate compound that, when combined withthe prostaglandin or retinoid, modulates the immune response to agreater degree than controls, is a compound that is potentially usefulfor treating a patient having an immunoinflammatory disorder or aproliferative skin disease.

Compounds useful in the invention include those described herein in anyof their pharmaceutically acceptable forms, including isomers such asdiastereomers and enantiomers, salts, solvates, and polymorphs thereof,as well as racemic mixtures of the compounds described herein.

The term “immunoinflammatory disorder” encompasses a variety ofconditions, including autoimmune diseases. Immunoinflammatory disordersresult in the destruction of healthy tissue by an inflammatory process.Examples of immunoinflammatory disorders include rheumatoid arthritis,ulcerative colitis, Crohn's disease, stroke-induced brain cell death,septic shock syndrome, ankylosing spondylitis, fibromyalgia, asthma,multiple sclerosis, type I diabetes, systemic lupus erythematosus,scleroderma, systemic sclerosis, inflammatory dermatoses, myastheniagravis, and Sjögren's syndrome. Inflammatory dermatoses include, forexample, psoriasis, acute febrile neutrophilic dermatosis, eczema (e.g.,asteatotic eczema, dyshidrotic eczema, vesicular palmoplantar eczema),balanitis circumscripta plasmacellularis, balanoposthitis, Behcetdisease, erythema annulare centrifugum, erythema dyschromicum perstans,erythema multiforme, granuloma annulare, lichen nitidus, lichen planus,lichen sclerosus et atrophicus, lichen simplex chronicus, lichenspinulosus, nummular dermatitis, pyoderma gangrenosum, sarcoidosis,subcorneal pustular dermatosis, urticaria, and transient acantholyticdermatosis.

The term “proliferative skin disease” encompasses benign and malignantproliferative skin diseases that are characterized by accelerated celldivision in the epidermis or dermis. Examples of proliferative skindiseases include psoriasis, atopic dermatitis, non-specific dermatitis,primary irritant contact dermatitis, allergic contact dermatitis, basaland squamous cell carcinomas of the skin, lamellar ichthyosis,epidermolytic hyperleratosis, premalignant keratosis, acne, andseborrheic dermatitis.

By “prostaglandin” is meant alprostidil, dinoprostone, misoprostil,prostaglandin E2, prostaglandin A1, prostaglandin A2, prostaglandin B1,prostaglandin B2, prostaglandin D2, prostaglandin Fla, prostaglandinF2α, prostaglandin I1, prostaglandin-ici 74205, prostaglandin F2β,6-keto-prostaglandin F1α, prostaglandin E1 ethyl ester, prostaglandin E1methyl ester, prostaglandin F2 methyl ester, arbaprostil, omoprostil,13,14-dihydroprostaglandin F2α, and prostaglandin J.

By “retinoid” is meant retinoic acid, retinol, and retinal, and naturalor synthetic derivatives of retinoic acid, retinol, or retinal that arecapable of binding to a retinoid receptor and consist of four isoprenoidunits joined in a head-to-tail manner. Examples of retinoids includetretinoin, vitamin A2 (3,4-didehydroretinol), α-vitamin A(4,5-didehydro-5,6-dihydroretinol), 13-cis-retinol, 13-cis retinoic acid(isotretinoin), 9-cis retinoic acid (9-cis-tretinoin), 4-hydroxyall-trans retinoic acid, torularodin, methyl retinoate, retinaldehyde,13-cis-retinal, etretinate, tazoretene, acetretin, alitretinoin andadapelene.

The combination of a prostaglandin and a retinoid for the treatment ofimmunoinflammatory disorders and proliferative skin diseases allows forthe administration of a low dose of each compound and less total activecompound, thus providing similar efficacy with less toxicity, andreduced costs.

Other features and advantages of the invention will be apparent from thefollowing detailed description, and from the claims.

DETAILED DESCRIPTION

We have discovered that the combination of the prostaglandin,alprostadil, with a retinoid, tretinoin, had substantial TNFαsuppressing activity on white blood cells. Concentrations thateffectively suppressed TNFα activity were not unacceptably toxic tonormal cells. Thus, combinations of prostaglandins and retinoids areuseful for the treatment of immunoinflammatory disorders andproliferative skin diseases.

Therapy

Combination therapy according to the invention may be performed alone orin conjunction with another therapy and may be provided at home, thedoctor's office, a clinic, a hospital's outpatient department, or ahospital. Treatment generally begins at a hospital so that the doctorcan observe the therapy's effects closely and make any adjustments thatare needed. The duration of the combination therapy depends on the typeof disease or disorder being treated, the age and condition of thepatient, the stage and type of the patient's disease, and how thepatient responds to the treatment. Additionally, a person having agreater risk of developing an immunoinflammatory disorder orproliferative skin disease (e.g., a person who is geneticallypredisposed or having a prior diagnosis of an immunoinflammatory orproliferative skin disorder) may receive prophylactic treatment toinhibit or delay the onset of symptoms.

A proliferative skin disease is alleviated when there is a noticeabledecrease in the size or thickness of a lesion to palpation. Thisdecrease can occur either with or without residual redness, dilatedblood vessels, hyper-pigmentation, or hypo-pigmentation. For thepurposes of this invention, psoriasis is considered alleviated when ascale-free psoriasis lesion is noticeably decreased in thickness.

The dosage, frequency and mode of administration of each component ofthe combination can be controlled independently. For example, onecompound may be administered orally three times per day, while thesecond compound may be administered intramuscularly once per day.Combination therapy may be given in on-and-off cycles that include restperiods so that the patient's body has a chance to recovery from any asyet unforeseen side-effects. The compounds may also be formulatedtogether such that one administration delivers both compounds.

Formulation of Pharmaceutical Compositions

Suitable modes of administration include oral, rectal, intravenous,intramuscular, subcutaneous, inhalation, topical or transdermal,vaginal, and ophthalmic. Administration of each compound of thecombination may be by any suitable means that results in a concentrationof the compound that, combined with the other compound, is effective.Each compound can be admixed with a suitable carrier substance, and isgenerally present in an amount of 1-95% by weight of the total weight ofthe composition. The pharmaceutical compositions may be formulatedaccording to conventional pharmaceutical practice (see, e.g., Remington:The Science and Practice of Pharmacy, (20th ed.) ed. A. R. Gennaro,2000, Lippencott Williams & Wilkens, Philadelphia, Pa., and Encyclopediaof Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan,1988-1999, Marcel Dekker, New York).

Pharmaceutical compositions according to the invention may be formulatedto release the active compound substantially immediately uponadministration or at any predetermined time period after administration,using controlled release formulations.

Administration of compounds in controlled release formulations is usefulwhere the compound, either alone or in combination, has (i) a narrowtherapeutic index (e.g., the difference between the plasma concentrationleading to harmful side effects or toxic reactions and the plasmaconcentration leading to a therapeutic effect is small; generally, thetherapeutic index, TI, is defined as the ratio of median lethal dose(LD₅₀) to median effective dose (ED₅₀)); (ii) a narrow absorption windowin the gastro-intestinal tract; or (iii) a short biological half-life,so that frequent dosing during a day is required in order to sustain theplasma level at a therapeutic level.

Many strategies can be pursued to obtain controlled release in which therate of release outweighs the rate of metabolism of the therapeuticcompound. For example, controlled release can be obtained by theappropriate selection of formulation parameters and ingredients,including, e.g., appropriate controlled release compositions andcoatings. Examples include single or multiple unit tablet or capsulecompositions, oil solutions, suspensions, emulsions, microcapsules,microspheres, nanoparticles, patches, and liposomes.

Topical Compositions

Therapeutic compositions suitable for topical application includeconventional anhydrous or aqueous preparations including ointments,lotions, creams, pastes, jellies, sprays, aerosols, and oils. Therepreparations can include oleaginous, aqueous, or emulsion-type bases.Optionally, topically applied formulations can be covered with anocclusive or semi-occlusive dressing.

Solid Dosage Forms for Oral Use

Formulations for oral use include tablets containing the activeingredient(s) in a mixture with non-toxic pharmaceutically acceptableexcipients. These excipients may be, for example, inert diluents orfillers (e.g., sucrose and sorbitol), lubricating agents, glidants, andantiadhesives (e.g., magnesium stearate, zinc stearate, stearic acid,silicas, hydrogenated vegetable oils, or talc).

The two compounds may be mixed together in a tablet or other vehicle, ormay be partitioned. In one example, the first compound is contained onthe inside of the tablet, and the second compound is on the outside,such that a substantial portion of the second compound is released priorto the release of the first compound.

Formulations for oral use may also be provided as chewable tablets, oras hard gelatin capsules wherein the active ingredient is mixed with aninert solid diluent, or as soft gelatin capsules wherein the activeingredient is mixed with water or an oil medium.

Dosages

The dosage of each compound of the claimed combinations used in anygiven therapeutic method depends on several factors, including: theadministration method, the condition to be treated, the severity of thecondition, whether the condition is to be treated or prevented, and theage, weight, and health of the person to be treated. Additionally,pharmacogenomic (the effect of genotype on the pharmacokinetic,pharmacodynamic or efficacy profile of a therapeutic) information abouta particular patient may affect dosage used.

As is described above, the compound(s) may be administered orally in theform of tablets, capsules, elixirs or syrups, or rectally in the form ofsuppositories. Parenteral administration of a compound is suitablyperformed, for example, in the form of saline solutions or with thecompound incorporated into liposomes. In cases where the compound initself is not sufficiently soluble to be dissolved, a solubilizer suchas ethanol can be applied. Below, the dosages for prostaglandins andretinoids are described.

For oral, intramuscular, subcutaneous, and intravenous administration ofthe prostaglandin, the dosage is normally about 1 pg to 10 mg per day,desirably about 10 pg to 1 mg per day, more desirably about 1 to 500 μgper day, and most desirably about 10 to 100 μg per day. Administrationcan be one to four times daily for one day to one year, and may even befor the life of the patient. Chronic, long-term administration will beindicated in many cases. For topical, inhalation, rectal, vaginal andophthalmic administration of the prostaglandin, the dosage is normallyabout 1 pg to 100 mg per day, desirably about 1 pg to 10 mg per day,more desirably about 100 pg to 1 mg per day, and most desirably about0.01 to 0.5 mg per day. Administration can be one to four times daily.Systemic dosing will result in steady-state plasma concentrations ofabout 1 picomolar to 1 micromolar, more desirably about 100 picomolar to100 nanomolar, and most desirably about 1 to 10 nanomolar.

For oral, intramuscular, subcutaneous, and intravenous administration ofthe retinoid, the dosage is about 1 μg to 5 g per day, desirably about0.1 mg to 1 g mg per day, more desirably about 1 to 100 mg per day, andmost desirably about 5 to 50 mg per day. For topical, inhalation,rectal, vaginal, or ophthalmic administration, the dosage is about 50 ngto 500 mg per day, desirably 500 ng to 50 mg per day, more desirablyabout 5 μg to 5 mg per day, and most desirably 50 to 500 μg per day.Administration can be one to four times daily for one day to one year,and may even be for the life of the patient. Chronic, long-termadministration will be indicated in many cases. Systemic dosing oftretinoin, for example, results in steady-state plasma concentrationdesirably of 500 picomolar to 50 micromolar, more desirably 5 nanomolarto 5 micromolar, and most desirably 50 to 500 nanomolar.

The following examples are to illustrate the invention. They are notmeant to limit the invention in any way.

EXAMPLE 1 Preparation of Combinations of Compounds

Stock solutions at 1.6 mg/ml of alprostadil, and 4.0 mg/ml of tretinoinacetate (Sigma-Aldrich, St. Louis, Mo.; catalog numbers P5515 andR-2625, respectively) were made in dimethylsulfoxide (DMSO). Thealprostadil master plates were made by adding 25 μl of the concentratedstock solution to columns 3, 9, and 15 (rows C through N) of apolypropylene 384-well storage plate that had been pre-filled with 75 μlof anhydrous DMSO. Using a TomTec Quadra Plus liquid handler, the 25 μlof alprostadil stock solution was serially diluted four times into theadjacent columns (columns 4-7, 10-13, 16-19). The sixth column (8, 14,and 20) did not receive any compound and served as a vehicle control.The tretinoin master plates were made by adding 25 μl of theconcentrated stock solution to the appropriate wells (row C, columns3-8; row C, columns 9-14; row C, columns 15-20; row I, columns 3-8; rowI, columns 9-14; row I, columns 15-20) of the appropriate masterpolypropylene 384-well storage plate. These master plates had beenpre-filled with 75 μl of anhydrous DMSO. Using the TomTec Quadra Plusliquid handler, the 25 μl was serially diluted four times in theadjacent rows (rows D-G, and J-M). The sixth row (H and N) did notreceive any compound to serve as a vehicle control. Master plates weresealed and stored at −20° C. until ready for use.

The final alprostadil/tretinoin combination plates were generated bytransferring 1 μl from each of the alprostadil and tretinoin masterplates to a dilution plate containing 100 μl of media (RPMI; Gibco BRL,#11875-085), 10% Fetal Bovine Serum (Gibco BRL, #25140-097), 2%Penicillin/Streptomycin (Gibco BRL, #15140-122)) using the TomTec QuadraPlus liquid handler. This dilution plate was then mixed and a 10 μlaliquot transferred to the final assay plate, which had been pre-filledwith 40 μl/well RPM media containing the appropriate stimulant toactivate TNFα secretion (see below).

EXAMPLE 2 Assay for TNFα Suppressing Activity by the Combination ofAlprostadil and Tretinoin

The compound dilution matrix was assayed using a TNFα ELISA method.Briefly, a 100 μl suspension of diluted human white blood cellscontained within each well of a polystyrene 384-well plate (NalgeNunc)was stimulated to secrete TNFα by treatment with a final concentrationof 10 ng/ml phorbol 12-myristate 13-acetate (Sigma) and 750 ng/mlionomycin (Sigma). Various concentrations of each test compound wereadded at the time of stimulation. After 16-18 hours of incubation at 37°C. in a humidified incubator, the plate was centrifuged and thesupernatant transferred to a white opaque polystyrene 384 well plate(NalgeNunc, Maxisorb) coated with an anti-TNFα antibody (PharMingen,#18631D). After a two-hour incubation, the plate was washed (TecanPowerWasher 384) with phosphate buffered saline (PBS) containing 0.1%Tween 20 (polyoxyethylene sorbitan monolaurate) and incubated for anadditional one hour with another anti-TNFα antibody that was biotinlabeled (PharMingen, 18642D) and horseradish peroxidase (HRP) coupled tostrepavidin (PharMingen, #13047E).). After the plate was washed with0.1% Tween 20/PBS, an HRP-luminescent substrate was added to each welland light intensity measured using a LJL Analyst plate luminometer. Setsof control wells contained a serial dilution of Cyclosporin A (Sigma)starting at a final concentration of 0.5 μg/ml.

Low doses of alprostadil significantly increased the ability oftretinoin to suppress TNFα secretion from stimulated white blood cells.As seen in Table 1, tretinoin alone maximally inhibited TNFα secretionby 36% at concentrations ranging from 0.83-13.3 μM. The combination of0.052 μM tretinoin and 0.004 μM alprostadil was able to suppress TNFαsecretion by 42%. The shift from 0.83 to 0.052 μM tretinoin without anyloss of activity represents a 16-fold increase in tretinoin potency inthe presence of low-dose alprostadil. Maximal tretinoin efficacy (−36%TNFα suppression) is therefore maintained while reducing theconcentration of total drug species by greater than 90% (0.052 μMtretionoin+0.004 μM alprostadil compared to 0.832 μM tretinoin alone).Table 1 also demonstrates that low-dose tretinoin potentiated TNFαsuppression by alprostadil. 1.130 M alprostadil inhibited TNFα secretionby 57%. The same level of inhibition was achieved by 0.071 μMalprostadil in the presence of 0.052 μM tretinoin. The 0.123 uMconcentration of the low-dose alprostadil/low-dose tretinoin combinationrepresents an 89% reduction in total drug species when compared to 1.130μM alprostadil alone.

Data from a secondary screen (Table 2) confirm and extend the observedsynergism between alprostadil and tretinoin. In this experiment, whiteblood cells were stimulated using 1 μg/ml LPS. TNFα inhibition bytretinoin alone was approximately 50%, at concentrations from 6.7-13.3μM. 59% inhibition of TNFα secretion was achieved by only 0.052 μMtretinoin in the presence of 0.004 μM alprostadil. The data also showthat the TNFα-suppressing activity of 0.208 μM tretinoin (32.5%) wasdoubled (64.0%) by the addition of 0.004 μM alprostadil. This level ofinhibition was not attainable by less than 0.035 μM alprostadil alone.Further evidence for the enhancement of alprostadil effects by tretinoinwere also measured. The maximal TNFα-inhibitory effect (82%) observedfor alprostadil alone required a concentration of at least 1.128 μM.This level of inhibition was exceeded (86%) by only 0.282 μM alprostadilin the presence of 0.104 μM tretinoin. TABLE 1 Alprostadil [μM]Tretinoin [μM] 1.130 0.283 0.071 0.018 0.004 0.000 13.316 80.62 79.5772.73 73.92 62.87 36.21 3.329 72.04 69.23 64.19 62.15 52.87 34.12 0.83270.58 68.71 65.20 57.69 54.71 34.53 0.208 68.35 65.45 59.56 54.48 48.8528.70 0.052 63.46 64.65 56.79 50.39 41.65 13.83 0.000 56.80 53.80 50.5239.75 33.30 0.00

TABLE 2 Alprostadil [μM] Tretinoin [μM] 1.128 0.564 0.282 0.141 0.0710.035 0.018 0.009 0.004 0.000 13.316 94.86 94.58 91.44 89.43 85.35 84.3279.81 76.18 72.63 52.08 6.658 93.50 91.23 88.53 88.56 84.81 78.32 68.7072.21 68.94 48.68 3.329 90.33 88.60 88.27 83.21 68.40 77.24 71.06 69.6063.24 44.10 1.664 89.37 89.34 78.92 84.76 77.01 75.18 73.03 67.84 67.1637.58 0.832 36.92 90.73 86.76 79.58 83.87 77.24 74.59 69.21 65.26 38.380.416 91.20 89.29 86.39 77.04 73.74 76.37 67.45 71.11 63.82 32.69 0.20887.47 90.44 88.16 83.41 70.91 73.53 62.12 69.30 64.01 32.50 0.104 85.0175.99 86.00 75.94 78.26 67.56 70.92 61.99 61.37 35.08 0.052 89.45 79.2278.62 73.82 75.04 69.51 68.82 65.83 58.84 32.71 0.000 81.60 79.45 75.6674.28 71.50 57.32 54.78 49.17 44.61 0.33

Other Embodiments

All publications and patents mentioned in the above specification areherein incorporated by reference. Various modifications and variationsof the described method and system of the invention will be apparent tothose skilled in the art without departing from the scope and spirit ofthe invention. Although the invention has been described in connectionwith specific preferred embodiments, it should be understood that theinvention as claimed should not be unduly limited to such specificembodiments. Indeed, various modifications of the described modes forcarrying out the invention that are obvious to those skilled in cellularand molecular biology, pharmacology, endocrinology, or related fieldsare intended to be within the scope of the invention.

1. A method for treating a patient who has an immunoinflammatorydisorder or a proliferative skin disease, or is at risk for developingan immunoinflammatory disorder or a proliferative skin disease, saidmethod comprising administering to said patient a prostaglandin and aretinoid, wherein the prostglandin and the retinoid are administeredsimultaneously or within 10 days of each other, in amounts sufficient totreat said patient.
 2. The method of claim 1, wherein said prostaglandinis alprostidil, misoprostil, dinoprostone, prostaglandin E2,prostaglandin A1, prostaglandin A2, prostaglandin B1, prostaglandin B2,prostaglandin D2, prostaglandin F1α, prostaglandin F2α, prostaglandinI1, prostaglandin-ici 74205, prostaglandin F2β, 6-keto-prostaglandinF1α, prostaglandin E1 ethyl ester, prostaglandin E1 methyl ester,prostaglandin F2 methyl ester, arbaprostil, omoprostil,13,14-dihydroprostaglandin F2α or prostaglandin J.
 3. The method ofclaim 2, wherein said prostaglandin is alprostadil or misoprostil. 4.The method of claim 1, wherein said retinoid is tretinoin, retinal,retinol, vitamin A2, α-vitamin A, 13-cis-retinol, isotretinoin,9-cis-tretinoin, 4-hydroxy all-trans retinoic acid, torularodin, methylretinoate, retinaldehyde, 13-cis-retinal, etretinate, tazoretene,acetretin, alitretinoin or adapelene.
 5. The method of claim 4, whereinsaid retinoid is tretinoin.
 6. The method of claim 1, wherein saidprostaglandin is alprostidil and said retinoid is tretinoin.
 7. Themethod of claim 1, wherein said prostaglandin and said retinoid areadministered within 5 days of each other.
 8. The method of claim 7,wherein said prostaglandin and said retinoid are administered within 24hours of each other.
 9. The method of claim 8, wherein saidprostaglandin and said retinoid are administered within one hour of eachother.
 10. The method of claim 9, wherein said prostaglandin and saidretinoid are administered simultaneously.
 11. The method of claim 1,wherein said immunoinflammatory disorder is rheumatoid arthritis,psoriasis, ulcerative colitis, Crohn's disease, an inflammatorydermatosis, septic shock syndrome, or stroke induced brain cell death.12. The method of claim 1, wherein said inflammatory dermatosis ispsoriasis.
 13. The method of claim 1, wherein said proliferative skindisease is contact dermatitis or acne.
 14. The method of claim 1,wherein said prostaglandin and said retinoid are administered to saidpatient by intravenous, intramuscular, subcutaneous, rectal, oral,topical, intravaginal, ophthalmic or inhalation administration.
 15. Themethod of claim 14, wherein said prostaglandin is administered in anamount of 1 pg to 100 mg per day, and said retinoid is administered inan amount of 50 ng to 5 g per day.
 16. The method of claim 15, whereinsaid prostaglandin is administered in an amount of 10 pg to 10 mg perday, and said retinoid is administered in an amount of 500 ng to 1 g perday.
 17. The method of claim 16, wherein said prostaglandin isadministered in an amount of 100 pg to 1 mg per day, and said retinoidis administered in an amount of 5 μg to 100 mg per day.
 18. The methodof claim 17, wherein said prostaglandin is administered in an amount of0.01 ng to 0.5 mg per day, and said retinoid is administered in anamount of 50 μg to 50 mg per day.
 19. A pharmaceutical compositioncomprising a prostaglandin, a retinoid, and a pharmaceuticallyacceptable carrier, wherein said prostaglandin and said retinoid areeach present in amounts that, when administered together to a patienthaving an immunoinflammatory disorder, inhibit or reduceimmunoinflammation or dermal/epidermal proliferation.
 20. Thepharmaceutical composition of claim 19, wherein said prostaglandin isalprostidil, misoprostil, dinoprostone, prostaglandin E2, prostaglandinA1, prostaglandin A2, prostaglandin B1, prostaglandin B2, prostaglandinD2, prostaglandin F1α, prostaglandin F2α, prostaglandin I1,prostaglandin-ici 74205, prostaglandin F2β, 6-keto-prostaglandin F1α,prostaglandin E1 ethyl ester, prostaglandin E1 methyl ester,prostaglandin F2 methyl ester, arbaprostil, omoprostil,13,14-dihydroprostaglandin F2α, or prostaglandin J.
 21. Thepharmaceutical composition of claim 20, wherein said prostaglandin isalprostidil or misoprostil.
 22. The pharmaceutical composition of claim19, wherein said retinoid is tretinoin, retinal, retinol, vitamin A2,α-vitamin A, 13-cis-retinol, isotretinoin, 9-cis-tretinoin, 4-hydroxyall-trans retinoic acid, torularodin, methyl retinoate, retinaldehyde,13-cis-retinal, etretinate, tazoretene, acetretin, alitretinoin oradapelene.
 23. The pharmaceutical composition of claim 22, wherein saidretinoid is tretinoin or retinol.
 24. The pharmaceutical composition ofclaim 19, wherein said prostaglandin is alprostidil and said retinoid istretinoin or retinol.
 25. The pharmaceutical composition of claim 19,wherein said prostaglandin and said retinoid are suitable forintravenous, intramuscular, subcutaneous, rectal, oral, topical,intravaginal, ophthalmic or inhalation administration.
 26. Apharmaceutical pack comprising a prostaglandin and a retinoid.
 27. Thepharmaceutical pack of claim 26, wherein said prostaglandin isalprostidil, misoprostil, dinoprostone, prostaglandin E2, prostaglandinA1, prostaglandin A2, prostaglandin B 1, prostaglandin B2, prostaglandinD2, prostaglandin F1α, prostaglandin F2α, prostaglandin I1,prostaglandin-ici 74205, prostaglandin F2β, 6-keto-prostaglandin F1α,prostaglandin E1 ethyl ester, prostaglandin E1 methyl ester,prostaglandin F2 methyl ester, arbaprostil, omoprostil,13,14-dihydroprostaglandin F2α, or prostaglandin J.
 28. Thepharmaceutical pack of claim 26, wherein said retinoid is tretinoin,retinal, retinol, vitamin A2, α-vitamin A, 13-cis-retinol, isotretinoin,9-cis-tretinoin, 4-hydroxy all-trans retinoic acid, torularodin, methylretinoate, retinaldehyde, 13-cis-retinal, etretinate, tazoretene,acetretin, alitretinoin or adapelene.
 29. The pharmaceutical pack ofclaim 26, wherein said prostaglandin and said retinoid are formulatedseparately and in individual dosage amounts.
 30. The pharmaceutical packof claim 26, wherein said prostaglandin and said retinoid are formulatedtogether and in individual dosage amounts.
 31. A method for identifyingcombinations of compounds useful for treating a patient having animmunoinflammatory disorder or proliferative skin disease, said methodcomprising the steps of: (a) contacting white blood cells in vitro with(i) a prostaglandin or a retinoid, and (ii); and a candidate compound;and (b) determining whether the combination of said prostaglandin orretinoid and said candidate compound reduces TNFα levels in said whiteblood cells relative to white blood cells contacted with saidprostaglandin or retinoid but not contacted with said candidatecompound, or white blood cells contacted with said candidate compoundbut not with said prostaglandin or retinoid, wherein a reduction of saidTNFα levels identifies said combination as a combination that is usefulfor treating a patient having an immunoinflammatory disorder orproliferative skin disease.